planar-gradient-diffusion-system-to-investigate-chemotaxis-3d

Planar Gradient Diffusion System to Investigate Chemotaxis in a 3D Collagen Matrix

The importance of cell migration can be seen through the development of human life. When cells migrate, they generate forces and transfer these forces to ...

Ximedica

Traction Force Microscopy (TFM) is a powerful approach for quantifying cell-material interactions that over the last two decades has contributed significantly to our understanding of cellular mechanosensing and mechanotransduction. In addition, recent advances in three-dimensional (3D) imaging and traction force analysis (3D TFM) have highlighted the significance of the third dimension in influencing various cellular processes. Yet irrespective of dimensionality, almost all TFM approaches have relied on a linear elastic theory framework to calculate cell surface tractions. Here we present a new high resolution 3D TFM algorithm which utilizes a large deformation formulation to quantify cellular displacement fields with unprecedented resolution. The results feature some of the first experimental evidence that cells are indeed capable of exerting large material deformations, which require the formulation of a new theoretical TFM framework to accurately calculate the traction forces. Based on our previous 3D TFM technique, we reformulate our approach to accurately account for large material deformation and quantitatively contrast and compare both linear and large deformation frameworks as a function of the applied cell deformation. Particular attention is paid in estimating the accuracy penalty associated with utilizing a traditional linear elastic approach in the presence of large deformation gradients.

High resolution, large deformation 3D traction force microscopy.

Native cell-material interactions occur on materials differing in their structural composition, chemistry, and physical compliance. While the last two decades have shown the importance of traction forces during cell-material interactions, they have been almost exclusively presented on purely elastic in vitro materials. Yet, most bodily tissue materials exhibit some level of viscoelasticity, which could play an important role in how cells sense and transduce tractions. To expand the realm of cell traction measurements and to encompass all materials from elastic to viscoelastic, this paper presents a general, and comprehensive approach for quantifying 3D cell tractions in viscoelastic materials. This methodology includes the experimental characterization of the time-dependent material properties for any viscoelastic material with the subsequent mathematical implementation of the determined material model into a 3D traction force microscopy (3D TFM) framework. Utilizing this new 3D viscoelastic TFM (3D VTFM) approach, we quantify the influence of viscosity on the overall material traction calculations and quantify the error associated with omitting time-dependent material effects, as is the case for all other TFM formulations. We anticipate that the 3D VTFM technique will open up new avenues of cell-material investigations on even more physiologically relevant time-dependent materials including collagen and fibrin gels.

3D Viscoelastic traction force microscopy.

Neutrophils are capable of switching from integrin-dependent motility on two-dimensional substrata to integrin-independent motion following entry into the confined three-dimensional matrix of an afflicted tissue. However, whether integrins still maintain a regulatory role for cell traction generation and cell locomotion under the physical confinement of the three-dimensional matrix is unknown, and this is challenging to deduce from motility studies alone. Using three-dimensional traction force microscopy and a double hydrogel sandwich system, we determined the three-dimensional spatiotemporal traction forces of motile neutrophils at unprecedented resolution and show, for the first time, that entry into a highly confined space (2.5D) is a sufficient trigger to convert to integrin-independent migration. We find that integrins exert a significant regulatory role in determining the magnitude and spatial distribution of tractions and cell speed on confined cells. We also find that 90% of neutrophil tractions are in the out-of-plane axis, and this may be a fundamental element of neutrophil traction force generation.

Jennet Toyjanova

Ximedica

Planar Gradient Diffusion System to Investigate Chemotaxis in a 3D Collagen Matrix